Nucleic acid sequence for a human beta tubulin

ABSTRACT

The present invention is related to a nucleic acid molecule comprising a nucleotide sequence encoding the C terminal end of a tubulin molecule, wherein the C terminal end comprises the following sequence:  
     VLEEDEEVTEEAEMEPEDKGH (SEQ ID. No. 4)

[0001] The present invention is related to nucleic acid moleculescomprising a nucleotide sequence encoding a tubulin molecule, vectorscontaining said nucleic acids, cells comprising such vectors, an aminoacid sequence coded by such nucleic acid molecule, use of such nucleicacids and methods for production of such tubulin molecules, fordiagnosing polymorphism in a tubulin molecule and for the screening ofanticancer drugs.

[0002] Microtubules are essential to the eucaryotic cell due as they areinvolved in many processes and functions such as, e.g., being componentsof the cytoskeleton, of the centrioles and ciliums and in the formationof spindle fibres during mitosis. The constituents of microtubules areheterodimers consisting of one α-tubulin molecule and one β-tubulinmolecule. These two related self-associating 50 kDa proteins are encodedby a multigen family. The various members of this multigen family aredispersed all over the human genome. Both α-tubulin and β-tubulin aremost likely to originate from a common ancestor as their amino acidsequence shows a homology of up to 50%. In man there are at least 15genes or pseudogenes for β-tubulin.

[0003] The conservation of structure and regulatory functions among theβ-tubulin genes in three vertebrate species (chicken, mouse and human)allowed the identification of and categorization into six major classesof beta-tubulin polypeptide isotypes on the basis of their variablecarboxyterminal ends. The specific, highly variable 15 carboxyterminalamino acids are very conserved among the various species. Beta-tubulinsof categories I, II, and IV are closely related differing only 2-4% incontrast to categories III, V and VI which differ in 8-16% of amino acidpositions [Sullivan K. F., 1988, Ann. Rev. Cell Biol. 4: 687-716].

[0004] Also the expression pattern is very similar between the variousspecies as can be taken from the following table [Sullivan K. F., 1988,Ann. Rev. Cell Biol. 4: 687-716] which comprises the respective humanmembers of each class: isotype member expression pattern class I HM 40ubiquitous class II H β 9 mostly in the brain class III H β 4exclusively in the brain class IVa H β 5 exclusively in the brain classIVb H β 2 ubiquitous

[0005] The C terminal end of the beta-tubulins starting from amino acid430 is regarded as highly variable between the various classes.Additionally, the members of the same class seem to be very conservedbetween the various species.

[0006] As tubulin molecules are involved in many processes and form partof many structures in the eucaryotic cell, they are possible targets forpharmaceutically active compounds. As tubulin is more particularly themain structural component of the microtubules it may act as point ofattack for anticancer drugs such as vinblastin, colchicin, estramustinand taxol which interfere with microtubule function. The mode of actionis such that cytostatic agents such as the ones mentioned above, bind tothe carboxyterminal end the β-tubulin which upon such binding undergoesa conformational change. For example, Kavallaris et al. [Kavallaris etal. 1997, J. Clin. Invest. 100: 1282-1293] reported a change in theexpression of of specific β-tubulin isotypes (class I, II, III, and IVa)in taxol resistant epithelial ovarian tumor. It was concluded that thesetubulins are involved in the formation of the taxol resistence. Also ahigh expression of class III β-tubulins was found in some forms of lungcancer suggesting that this isotype may be used as a diagnostic marker.

[0007] The problem underlying the present invention was to provide themeans to further characterize the various tubulins present in eucaryoticcells. A further problem underlying the present invention was to providethe means to extend possible screening programs for cytostatic agents toother isotypes of human beta-tubulins.

[0008] This problem is solved in a first aspect by a nucleic acidmolecule comprising a nucleotide sequence encoding a tubulin molecule,wherein said nucleic acid molecule comprises the sequence according toSEQ. ID. No. 1

[0009] This problem is solved in a second aspect by a nucleic acidmolecule comprising a nucleotide sequence encoding a tubulin molecule,wherein said nucleic acid molecule comprises the sequence according toSEQ. ID. No. 2.

[0010] This problem is solved in a third aspect by a nucleic acidmolecule comprising a nucleotide sequence encoding the C terminal end ofa tubulin molecule, wherein the C terminal end comprises the followingsequence:

[0011] VLEEDEEVTEEAEMEPEDKGH (SEQ ID. No. 4)

[0012] This problem is solved in a fourth aspect by a nucleic acidmolecule comprising a nucleotide sequence encoding a tubulin molecule,wherein said nucleic acid molecule hybridises to the nucleic acidmolecule selected from the group comprising SEQ. ID. No. 1, SEQ. ID. No2, SEQ. ID. No. 4 and derivatives therefrom.

[0013] This problem is solved in a fifth aspect by a nucleic acidmolecule comprising a nucleotide sequence encoding a tubulin molecule,wherein the tubulin molecule or part thereof comprises an amino acidsequence according to SEQ. ID. No. 3.

[0014] This problem is solved in a sixth aspect by a nucleic acidmolecule comprising a nucleotide sequence encoding a tubulin molecule,wherein the nucleotide nucleic acid molecule would hybridise, but forthe degeneracy of the genetic code, to any of the nucleic acid moleculeand/or nucleotide sequence according to the above aspects of theinvention.

[0015] In a further aspect the problem is solved by an amino acidsequence of a tubulin molecule, preferably of a beta-tubulin moleculeand more preferably of a class VI beta-tubulin, wherein the amino acidsequence is coded by a nucleic acid molecule according to the presentinvention or part thereof.

[0016] In a preferred embodiment of the various aspects of the inventionthe tubulin molecule is the tubulin molecule expressed in haematopoietictissue, in an even more preferred embodiment the tubulin molecule ispreferably expressed in haematopoietic tissue.

[0017] In a further preferred embodiment of the various aspects of theinvention the tubulin molecule is β-tubulin, preferably an isotype classVI beta-tubulin, most preferably a human homologue of the vertebratebeta tubulin isotype class VI.

[0018] In a still further preferred embodiment of the various aspects ofthe invention the nucleic acid molecule is an isolated nucleic acidmolecule.

[0019] In a seventh aspect of the invention the problem is solved by avector comprising any of the nucleic acid molecules of the presentinvention.

[0020] In a preferred embodiment the vector is an expression vector.

[0021] In a eighth aspect of the invention the problem is solved by acell, preferably a host cell, comprising the vector according to theinvention.

[0022] In a nineth aspect of the invention the problem is solved by amethod for the production of a tubulin molecule, preferably of a beta-tubulin molecule and more preferably of a class VI beta-tubulin,characterized by culturing the cell according to the present inventionand isolating the protein product.

[0023] In a tenth aspect of the invention the problem is solved by amethod for diagnosing polymorphism in a tubulin molecule or gene,preferably β-tubulin, and more preferably an isotype class VIbeta-tubulin, most preferably a human homologue of the vertebrate betatubulin isotype class VI or in the nucleic acid coding such tubulin,wherein a nucleic acid according to the present invention is used.

[0024] In a eleventh aspect of the invention the problem is solved bythe use of a nucleic acid according to the present invention fordiagnosis.

[0025] In a twelfth aspect of the invention the problem is solved by amethod for the screening for anticancer drugs using a nucleic acidaccording to any of claims 1 to 9 or any transcription and/ortranslation product thereof.

[0026] In a further aspect of the invention the problem is solved by theuse of a nucleic acid according to the present invention for screeningpurposes.

[0027] The inventors of the present invention have surprisingly foundthe human homologue of the vertebrate beta-tubulin isotype class VI.This homologue can be characterised by the C-terminal amino acidsequence. A comparison of the C-terminal isotype defining sequencesincluding the one of the human homologue referred to herein also asTUBB1, can be taken from table 1. TABLE 1 C-terminal isotype definingsequences. C-terminal Isotype isotype defining sequence gene/proteinClass I EEEEDFGEEAEEEA cb7 Class II DEQGEFEEEGEEDEA cb1/cb2          EGmb2 Class III EEEGEMYEDDEEESEQGAK cb4                   P hb4 Class IVaEEGEFEEEAEEEVA mb4 Class IVb EEEGEFEEEAEEEAE cb3              VA mb3Class V NDGEEEAFEDDEEEINE cb5 Class VI DVEEYEEAEASPEKET cb6GLEDSEEDAEEAEVEAEDKDH mb1 V  ED  VT    M P  G TUBB1

[0028] The remainder of the sequences are identical. Particularlyfocussing on the C-terminal isotype defining sequences it is to be notedthat the class VI isotypes are so variable that the sequences of chicken(cb6) and mouse (mb1) are completely specified whereas TUBB1 is ratherhomologue so that the only differences of TUBB1 relative to mb1 areindicated. In connection with the beta-tubulin isotypes of table 1 <<h>>refers to human, <<c>> to chicken and <<m>>to mouse>>. The h homologueTUBB1 shows a homology of 92% relative to mb1 on the basis of the 451 AStranslated out of the potential ORF.

[0029] Derivatives of the various nucleic acids described herein areparticularly those nucleic acids which encode the same amino acids asdoes the non-derivative nucleic acid, or those nucleic acids which showthe same chemical, biological or structural characteristics as does thenon-derivative one. Such derivative typically comprises at least somenucleobases which differ from the nucleobases of the non-derivativenuleic acids. Preferably a pyrimidine nucleobase is replaced by anotherpyrimidine nucleobase and a purine nucleobase replaced by a differentpurine nucleobase. Derivatives of a nucleic acid in the meaning of thisinvention may also be such nucleic acid (and amino acids possiblyderived therefrom) which hybridise to said nucleic acid. Moreparticularly, such hybridisation is carried out using an oligonucleotidecomprising the following sequence (SEQ ID No. 14): GTT CTA GAG GAA GATGAA GAG GTC ACG GAG GAG GCA GAA ATG GAG CCA GAA GAT AAG GGA CAT

[0030] As probe for stringent hybridisation meaning a hybridisationsolution with a Na+ concentration of 0.4 M and a hybridisationtemperature range of 55 to 60° C. SEQ. ID. No. 14 is based on SEQ ID.No. 4

[0031] The C-terminal sequence of TUBB1 is the part of the beta-tubulinwhere MAPs (microtubuli associated proteins) preferably bind which areproteins partially unknown having an impact on association ofmicrotubuli. Similar to the mouse homologue mb1 (Wang et. al 1986, J.Cell Biol. 103: 1903-1910) and its translation product (Kavallaris etal. 1997, J. Clin. Invest. 100: 1282-1293) the TUBB1 protein is found inthe cells of the hematopoietic tissue as shown for mb1 in bone marrowand thrombocytes by Western blot analysis. It is understood that theprotein of isotype VI is specific for the Marginal Band in nucleatedfetal erythroblasts and platelets, respectively (Lewis et al. 1987, Cell49 (4): 539-48)

[0032] Having the sequence of the human homologue of beta-tubulinisotype class VI now at hand, various applications thereof are possible.One such application is the screening for cytostatic drugs. Basically,one such approach is the same as described by Kavallaris (supra) thedisclosure of which is herein incorporated by reference, and is based onthe interference of a candidate cytostatic drug with the aggregation ofthe tubulin molecules. Such screening can be carried out in cellculture.

[0033] A further application is the use of the TUBB1 and its amino acidand nucleic acid sequence, respectively, for the diagnosis ofpolymorphic regions within the TUBB1 gene and for a predisposition fordistinct diseases such as leucaemia and other diseases related tohaematopoiesis.

[0034] Vectors useful in the practice of the present invention are knownto the one skilled in the art, particularly those which provide for afurther method for purification of the expressed protein such aspPROTet.E/pPROLar.A (Clontech). With regard to a possibleposttranslational processing an expression in a baculovirus system isalso envisaged which means that vectors such as, e.g., pBacPAK 8/9(Clontech) may be used. Inducible vector systems may also be used,preferably for testing the effect of TUBB1 in a cellular environment.Such testing may be part of a test for therapeutical approaches.Representative vectors and cellular systems are pRevTRE (Clontech) andHeLa (epithelial carcinoma, cervix). Other suitable cell lines useful inthe practice of the present invention are known to the one skilled inthe art.

[0035] Further molecular biological methods useful in the practice ofthe present invention are, e.g., described in Maniatis et al. (ManiatisT., Fritsch E. F., Sambrook J., (1982) Cold Spring Harbor Press, ColdSpring Harbor).

[0036] The invention is now further characterized by reference to thesequences of the sequence protocol, the figures and the examples fromwhich further embodiments, features and advantages can be taken, wherein

[0037] SEQ. ID. No. 1 is the cDNA sequence of TUBB1;

[0038] SEQ. ID. No. 2 is the open reading frame of the cDNA;

[0039] SEQ. ID. No.3 is the amino acid sequence of the open readingframe of SEQ. ID. No. 2;

[0040] SEQ. ID. No. 4 is the C-terminal isotype identifying sequence ofthe human homologue of the beta-tubulin isotype class VI;

[0041]FIG. 1 shows a 5′RACE for H57679 using primers AP2/H57E3F (1) andAP2/H57E1F (3);

[0042]FIG. 2 shows subclones of J0464 for the sequencing; and

[0043]FIG. 3 shows the arrangement of the cDNA clones for H57679.

[0044]FIG. 1 shows the 5′RACE for H57679 out of a marathon-ready-cDNAlibrary. Lane 1 shows a 260 bp fragment amplified during PCR usingsequence specific primer H57E2F and adaptor specific primer AP2 whereaslane 2 shows similar fragments using different sequence specific primerH57E1F and AP2. Lanes 3 and 4 are control PCR to exclude anycontamination.

[0045] Sequencing of the 3.3 kb insert of J0464 is based on producingseveral subclones using the Genome Priming System (NEB, New EnglandBiolabs, Schwalbach, Germany). The statistical integration of aTransposon element allows a sequence specific selection of positiveclones using PCR as shown in FIG. 2. PCR analysis of 20 subclones usingsequence specific primer H57679L and the Transposon specific primer Tn7Rshows several different integration sides which could be used forsequencing of overlapping fragments. H57679L: tgactggcatcctgctttc (SEQ.ID. No. 5) Tn7R: gcataactggactgatttc (SEQ ID. No. 6)

EXAMPLE 1 Characterisation of TUBB1

[0046] The gene of SEQ. ID. No. 1, referred to herein as TUBB1, wascharacterized starting from an EST in the distal part of chromosome 20 qwhich was taken from the Radiation Hybrid Map (RH4) which is a databasebased physical map of the human genome. The Radiation Hybrid Database byPatricia Rodriguez-To{acute over (m)}, Carsten Helgesen and PhilipLijnzaad can be characterised as follows: The EBI (EMBL Outstation, TheEuropean Bioinformatics Institute, Wellcome Trust Genome Campus,Hinxton, Cambridge CB10 1SD,UK; since July 1995) maintains RHdb, apublic database for radiation hybrid mapping, including raw results andmaps. RH data are used in the generation of alternative genetic maps asthey can include non-polymorphic markers. The main type of entry in thedatabase consists of PCR hybridization results of a particular STS on aparticular panel. STS belonging to one (or more) of the followingcategories are used for mapping:

[0047] Genetic markers

[0048] Expressed Sequences Tags (EST)

[0049] Fully sequenced cDNA

[0050] CpG Islands

[0051] Random genomic human clones

[0052] Chromosome specific libraries.

[0053] Cross-references to a number of related databases are alsostored. These include Genbank/EMBL, dbEST, GDB, and the databases of thelaboratories that did the experiments. The database is maintained in theOracle Relational Database Management System (RDBMS).

[0054] The Radiation Hybrid Map itself was taken from the website ofNCBI (National Center for Biotechnology Information (NCBI); NationalCenter for Biotechnology Information, National Library of Medicine,Building 38A, Room 8N805, Bethesda, Md. 20894). The particular ESTchosen is H 57679. EST 57679 is one out of many ESTs located in thedistal area of chromosome 20. The entry of the EMBL database reads asfollows:

[0055] LOCUS H57679 433 bp mRNA EST 05, Oct. 1995

[0056] DEFINITION yr27b11.s1 Soares fetal liver spleen 1NFLS Homosapiens cDNA clone

[0057] IMAGE:206493 3′, MRNA sequence.

[0058] ACCESSION H57679

[0059] VERSION H57679.1 GI:1010511

[0060] KEYWORDS EST.

[0061] SOURCE human.

[0062] ORGANISM Homo sapiens Eukaryota; Metazoa; Chordata; Craniata;Vertebrata; Euteleostomi; Mammalia; Eutheria; Primates; Catarrhini;Hominidae; Homo.

[0063] REFERENCE 1 (bases 1 to 433)

[0064] AUTHORS Hillier, L., Clark, N., Dubuque, T., Elliston, K.,Hawkins, M., Holman, M., Hultman, M., Kucaba,T., Le, M., Lennon, G.,Marra, M., Parsons, J., Rifkin, L., Rohlfing, T., Soares, M., Tan, F.,Trevaskis, E., Waterston, R., Williamson, A., Wohldmann, P. and Wilson,R.

[0065] TITLE The WashU-Merck EST Project

[0066] JOURNAL Unpublished

[0067] COMMENT Contact: Wilson RK Washington University School ofMedicine, 4444 Forest Park Parkway, Box 8501, St. Louis, Mo. 63108.

[0068] The cDNA clone corresponding to EST H 57679 was ordered from theRZPD in Berlin, Germany (Resource Center/Primary Database: Heubnerweg 6,14059 Berlin)

[0069] After sequencing of the cDNA clone using standard primers(M13rev, -21M13for, T3, T7), respectively, having the followingsequences: −21M13forward: TGTAAAACGACGGCCAGT (SEQ. ID. No. 7) andM13reverse: CAGGAAACAGCTATGACC, (SEQ. ID. No. 8) T3:AATTAACCCTCACTAAAGGG; (SEQ. ID. No. 9) T7: TAATACGACTCACTATAGGG, (SEQ.ID. NO. 10)

[0070] primers were derived therefrom and used for the screening offurther cDNA clones from tissue specific cDNA libraries of the RZPD(multiple tissue pools; MTP) which are also referred to as “GeneFinder”pools.

[0071] The following further clones were found: name of the clone seizeof the insert ICRFp512B12195Q1 1.7 kb ICRFp5I2J0464Q1 3.3 kbICRFp512J12195Q1 1.7 kb ICRFp512L14188Q1 1.3 kb

[0072] For further verification of the completeness of the 5′end of thecDNA a RACE-PCR from Marathon-Ready-cDNA (CLONTECH GmbH, Tullastr. 4,69126 Heidelberg, Germany) was run using the following primers: AP2actcactatagggctcgagcggc (SEQ. ID. No. 11) H57E1F cacggaatcctcagactctcag(SEQ. ID. No. 12) H57E2F gttgtagtacacgctgattctctcc (SEQ. ID. No. 13)

[0073] AP2 is an adaptor specific primer. The primers H57 E1F and H57E2Fare specific for sequences of the 5′part of clone ICRFp512J0464Q1.

[0074] The details of the RACE-PCR can be summarized as follows: In afirst round an adaptor specific and a gene specific primer are used; inthe second round a re-amplification is carried our using the sameprimers. 1st round: 10 pg/μl Marathon-ready-cDNA 1x cDNA PCR-reactionbuffer (Clontech) 0.2 mM dNTPs 1x Advantage cDNA Pol. mix 0.2 pmolAdaptor primer 0.2 pmol H57E2F ad 5 μl H₂O 1:50 out of 1. 1x Elongasebuffer (Gibco BRL) 0.5 mM dNTPs 1.95 mM MgCl₂ 0.25 μl Elongase (GibcoBRL) 0.2 pmol Adaptor primer 0.2 pmol H57E2F ad 10 μl H₂O PCR: 22x 92°C. - 50 s 72° C. - 50 s (−1° C. per cycle) 68° C. - 4 min 25x 92° C. -50 s 60° C. - 50 s 68° C. - 4 min

[0075] A PCR product of about 260 bp was amplified and directlysequenced by using the primers H57E2F/AP2 applied in the RACE-PCR.

[0076] Analysis of the results indicated that the 5′ end ofICRFp512J0464Q1 was to be regarded as complete as the sequence of theMarathon product was identical except four bases which confirmed thesequence of the clone. As the 3 ′ends of all clones were nearlyidentical there is now a complete or full-length cDNA for thislocus/gene on chromosome 20q.

EXAMPLE 2

[0077] Sequences homologous to TUBB1

[0078] With regard to the fact that typically isotypes of one class oftubulins share a high degree of homology between various species arespective analysis of class VI isotype beta-tubulins was performed onhomology at the amino acid level the results of which are summarized inthe following table. The basis for this analysis is the amino acidsequence of TUBB1 corresponding to SEQ. ID. No. 3. Table Acc. Nr. Ident.Id. Pos. Pos. (%) (%) Mouse beta tubulin mb1 413/451 438/451 92 97Chicken class VI P09207 353/430 389/430 82 90 beta tubulin

[0079] Both mouse beta tubulin mb1 and chicken class VI beta tubulin aredescribed in literature. Chicken: Murphy D. B., Wallis K. T., Machlin P.S., Ratrie H. III, Cleveland D. W.; “The sequence and expression of thedivergent β-tubulin in chicken erythrocytes.”; J. Biol. Chem.262:14305-14312(1987)). The accession number refers to SWISSPROT; thecDNA is also available from EMBL under accession number J02828.

[0080] The mouse sequence is available neither from EMBL nor fromSWISSPROT, but was published by Wang et al. (Wang D, Villasante A, LewisS A, Cowan N J “The mammalian beta-tubulin repertoire: hematopoieticexpression of a novel, heterologous beta-tubulin isotype” J. Cell Biol.103(5):1903-10 (1986)).

[0081] The features disclosed in the specification, the figures,sequence protocol and the claims may be, individually or in combination,essential for the realization of the invention in its variousembodiments.

1 14 1 3495 DNA Homo sapiens 1 ggggcagtat tctgtgttga gggaggaaaaacactccctt ccaaaagcat gacaggcaga 60 aagcagagaa gggccaggac tggctgagggcggggagctg ggcctctggg gtggacacac 120 ccttggtcac attgtgaggg tagcttggttggccagtccc accactgcag tgaccacagt 180 tgtgttgggc tcacaccagt gaaccgaagctctggattct gagagtctga ggattccgtg 240 aagatctcag acttgggctc agagcaaggatgcgtgaaat tgtccatatt cagattggcc 300 agtgtggcaa ccagatcgga gccaagttctgggagatgat tggtgaggaa cacgggatcg 360 acttggctgg gagcgaccgc ggggcctcggccttgcagct ggagagaatc agcgtgtact 420 acaacgaagc ctacggtagg aaatatgtgccccgagcagt cttggtggac ctagaacctg 480 ggacgatgga cagcattcga tctagcaaattaggagctct ctttcaaccc gacagttttg 540 tccatggtaa ctctggggct ggcaacaactgggccaaagg ccactacacg gagggagccg 600 agctgatcga gaatgtccta gaggtggtgaggcacgagag tgagagctgt gactgcctgc 660 agggcttcca gatcgtccac tccctgggcgggggcacagg ctccgggatg ggcactctgc 720 tcatgaacaa gattagagag gagtacccggaccggatcat gaattccttc agcgtcatgc 780 cttctcccaa ggtgtcggac acggtggtggagccctacaa cgcggttctg tctatccacc 840 agctgattga gaatgcagat gcctgtttctgcattgacaa tgaggccctc tatgacatct 900 gcttccgtac cctgaagctg acgacacccacctatgggga tctcaaccac ctagtgtcct 960 tgaccatgag cggcataacc acctccctccggttcccggg tcagctcaac gcagacctgc 1020 gcaagctggc ggtgaacatg gtccccttcccccgcctgca cttctttatg cccggctttg 1080 ccccactcac ggcccagggc agccagcagtaccgagccct ctccgtggcc gagctcaccc 1140 agcagatgtt cgatgcccgc aataccatggctgcctgtga cctccgccgt ggccgctacc 1200 tcacagtggc ctgcattttc cggggcaagatgtccaccaa ggaagtggac cagcaactgc 1260 tctccgtgca gaccaggaac agcagctgctttgtggagtg gattcccaac aacgtcaagg 1320 tggctgtctg cgacatcccg ccccgggggctgagcatggc cgccaccttc attggcaaca 1380 acacggccat ccaagagatc tttaatagggtctctgagca tttctcagcc atgttcaaaa 1440 ggaaagcttt tgtgcactgg tacaccagcgaagggatgga cataaacgaa tttggggaag 1500 ctgaaaataa catccatgat ttggtatccgagtaccaaca atttcaagat gccaaagcag 1560 ttctagagga agatgaagag gtcacggaggaggcagaaat ggagccagaa gataagggac 1620 attaactgtg agagaagctg tgccgcggagtcgcttacag aacagtttct cattagatga 1680 gtgtttctcc tgcagcactc caaaacccactctgcactgc agcacagtga atgatatgca 1740 ctcaccatta gcttcgacac agggactgagggagacaggt ggggagcagc tgacaggcat 1800 tagggtcttt gctgacatct actaaccttgaagagtttga tgttcagtgc atacttatta 1860 acttaaaaaa atagcaaatt tattgtaaagtgctcccttt gtttcaaagt gtttgccagg 1920 catccagact acacgtgtgg atttgcagggagccactgga gttggtgtta catttttata 1980 ctttagcagc actgataggc accctggaatcctcacttgg tatccgaggg ctactaagac 2040 tctttcctta ggttctttcc tctgagcaaacactgactgg catcctgctt tccagtgcct 2100 gccagcctcc agaagagcca ggtgcctgactagtacatgg ggagctacag agccaaggtc 2160 aatgtgagtc aacatccact agaaatatccatgttgtgta gacctgtgca tacaacatgc 2220 taactggaaa agaggaaaaa agaaaagccacagtcctctc cacaaaaata cctggtccaa 2280 acaagaaaaa caaaaagaca agcaaaactaaagaactgca gtcttctgat ctttatttct 2340 gaagagctag cctttaacat atatgtttatatagtttaaa tttcttacta ctgttagatc 2400 ccaggaattc attaataatc atccttggctttccttttaa aggctatttt gaaatggtct 2460 tttcactttc attcagtcat caccccccaaaatgctctgc agcctctctg ctctttgaga 2520 aagggcacac catgcgctcg gcaaccattcaaatgcagga attaagcagc aatggctgca 2580 gtgtccttct cagttatgga ggacatcgtctcattaggga acttttacag ttcaaattaa 2640 tttgcagaag ttgccataaa tgtttgcataatgacatagc tttagcacta catgatttta 2700 atctgctcac attataacag gaccaaatacacaagagcgt aatcaaatca tctgtaactt 2760 cttaattaca gtttacctat ttctgacatgcagcactgcc atctcttcca gcaccatcag 2820 ggttttaatg gccctctaga attaccactgagatacacta tttgatccat ggataaccgg 2880 taatgggaaa atgctccgac cctcaatgcagtaaatattt acttgcaggc aactgggttc 2940 tcatctcttg atttgctttt gtaatcagcaataataaaat agcaggtaga tggatgacag 3000 ttgctcattc tgagaaactt cactcttttcacttatgcat cacgaggaaa taactaaaat 3060 acataccaag agaaaaatac cttgccatcggatcatcaac aagtcttcta tttacaaact 3120 tcaaaaaaca caaaacaaca ttcatgttttaaatgctttc tacttgtggt tcaagaagca 3180 ctagatttag taagaaactc tacctatatacttagtttga agttagtaac ttcctgagat 3240 gctaaagact tacagcctgc gattatacaaggatttacac atgcttcctc tggtgcttta 3300 cttcccaaac ctaaaaaagc aatgaaatagatgtaaggaa ggagggattt aaacctttta 3360 aaaaactttt gctgacttat attactgtaaagatttgttt gctcaatagt aatcattaaa 3420 ctacaaagta attcaatttt aaatggcaaaattgctttat ttcagactaa ataaattcct 3480 tttcttgaag cctaa 3495 2 1356 DNAHomo sapiens 2 atgcgtgaaa ttgtccatat tcagattggc cagtgtggca accagatcggagccaagttc 60 tgggagatga ttggtgagga acacgggatc gacttggctg ggagcgaccgcggggcctcg 120 gccttgcagc tggagagaat cagcgtgtac tacaacgaag cctacggtaggaaatatgtg 180 ccccgagcag tcttggtgga cctagaacct gggacgatgg acagcattcgatctagcaaa 240 ttaggagctc tctttcaacc cgacagtttt gtccatggta actctggggctggcaacaac 300 tgggccaaag gccactacac ggagggagcc gagctgatcg agaatgtcctagaggtggtg 360 aggcacgaga gtgagagctg tgactgcctg cagggcttcc agatcgtccactccctgggc 420 gggggcacag gctccgggat gggcactctg ctcatgaaca agattagagaggagtacccg 480 gaccggatca tgaattcctt cagcgtcatg ccttctccca aggtgtcggacacggtggtg 540 gagccctaca acgcggttct gtctatccac cagctgattg agaatgcagatgcctgtttc 600 tgcattgaca atgaggccct ctatgacatc tgcttccgta ccctgaagctgacgacaccc 660 acctatgggg atctcaacca cctagtgtcc ttgaccatga gcggcataaccacctccctc 720 cggttcccgg gtcagctcaa cgcagacctg cgcaagctgg cggtgaacatggtccccttc 780 ccccgcctgc acttctttat gcccggcttt gccccactca cggcccagggcagccagcag 840 taccgagccc tctccgtggc cgagctcacc cagcagatgt tcgatgcccgcaataccatg 900 gctgcctgtg acctccgccg tggccgctac ctcacagtgg cctgcattttccggggcaag 960 atgtccacca aggaagtgga ccagcaactg ctctccgtgc agaccaggaacagcagctgc 1020 tttgtggagt ggattcccaa caacgtcaag gtggctgtct gcgacatcccgccccggggg 1080 ctgagcatgg ccgccacctt cattggcaac aacacggcca tccaagagatctttaatagg 1140 gtctctgagc atttctcagc catgttcaaa aggaaagctt ttgtgcactggtacaccagc 1200 gaagggatgg acataaacga atttggggaa gctgaaaata acatccatgatttggtatcc 1260 gagtaccaac aatttcaaga tgccaaagca gttctagagg aagatgaagaggtcacggag 1320 gaggcagaaa tggagccaga agataaggga cattaa 1356 3 451 PRTHomo sapiens 3 Met Arg Glu Ile Val His Ile Gln Ile Gly Gln Cys Gly AsnGln Ile 1 5 10 15 Gly Ala Lys Phe Trp Glu Met Ile Gly Glu Glu His GlyIle Asp Leu 20 25 30 Ala Gly Ser Asp Arg Gly Ala Ser Ala Leu Gln Leu GluArg Ile Ser 35 40 45 Val Tyr Tyr Asn Glu Ala Tyr Gly Arg Lys Tyr Val ProArg Ala Val 50 55 60 Leu Val Asp Leu Glu Pro Gly Thr Met Asp Ser Ile ArgSer Ser Lys 65 70 75 80 Leu Gly Ala Leu Phe Gln Pro Asp Ser Phe Val HisGly Asn Ser Gly 85 90 95 Ala Gly Asn Asn Trp Ala Lys Gly His Tyr Thr GluGly Ala Glu Leu 100 105 110 Ile Glu Asn Val Leu Glu Val Val Arg His GluSer Glu Ser Cys Asp 115 120 125 Cys Leu Gln Gly Phe Gln Ile Val His SerLeu Gly Gly Gly Thr Gly 130 135 140 Ser Gly Met Gly Thr Leu Leu Met AsnLys Ile Arg Glu Glu Tyr Pro 145 150 155 160 Asp Arg Ile Met Asn Ser PheSer Val Met Pro Ser Pro Lys Val Ser 165 170 175 Asp Thr Val Val Glu ProTyr Asn Ala Val Leu Ser Ile His Gln Leu 180 185 190 Ile Glu Asn Ala AspAla Cys Phe Cys Ile Asp Asn Glu Ala Leu Tyr 195 200 205 Asp Ile Cys PheArg Thr Leu Lys Leu Thr Thr Pro Thr Tyr Gly Asp 210 215 220 Leu Asn HisLeu Val Ser Leu Thr Met Ser Gly Ile Thr Thr Ser Leu 225 230 235 240 ArgPhe Pro Gly Gln Leu Asn Ala Asp Leu Arg Lys Leu Ala Val Asn 245 250 255Met Val Pro Phe Pro Arg Leu His Phe Phe Met Pro Gly Phe Ala Pro 260 265270 Leu Thr Ala Gln Gly Ser Gln Gln Tyr Arg Ala Leu Ser Val Ala Glu 275280 285 Leu Thr Gln Gln Met Phe Asp Ala Arg Asn Thr Met Ala Ala Cys Asp290 295 300 Leu Arg Arg Gly Arg Tyr Leu Thr Val Ala Cys Ile Phe Arg GlyLys 305 310 315 320 Met Ser Thr Lys Glu Val Asp Gln Gln Leu Leu Ser ValGln Thr Arg 325 330 335 Asn Ser Ser Cys Phe Val Glu Trp Ile Pro Asn AsnVal Lys Val Ala 340 345 350 Val Cys Asp Ile Pro Pro Arg Gly Leu Ser MetAla Ala Thr Phe Ile 355 360 365 Gly Asn Asn Thr Ala Ile Gln Glu Ile PheAsn Arg Val Ser Glu His 370 375 380 Phe Ser Ala Met Phe Lys Arg Lys AlaPhe Val His Trp Tyr Thr Ser 385 390 395 400 Glu Gly Met Asp Ile Asn GluPhe Gly Glu Ala Glu Asn Asn Ile His 405 410 415 Asp Leu Val Ser Glu TyrGln Gln Phe Gln Asp Ala Lys Ala Val Leu 420 425 430 Glu Glu Asp Glu GluVal Thr Glu Glu Ala Glu Met Glu Pro Glu Asp 435 440 445 Lys Gly His 4504 21 PRT Artificial Sequence Description of Artificial SequencePrimer 4Val Leu Glu Glu Asp Glu Glu Val Thr Glu Glu Ala Glu Met Glu Pro 1 5 1015 Glu Asp Lys Gly His 20 5 19 DNA Artificial Sequence Description ofArtificial SequencePrimer 5 tgactggcat cctgctttc 19 6 19 DNA ArtificialSequence Description of Artificial SequencePrimer 6 gcataactgg actgatttc19 7 18 DNA Artificial Sequence Description of Artificial SequencePrimer7 tgtaaaacga cggccagt 18 8 18 DNA Artificial Sequence Description ofArtificial SequencePrimer 8 caggaaacag ctatgacc 18 9 20 DNA ArtificialSequence Description of Artificial SequencePrimer 9 aattaaccctcactaaaggg 20 10 20 DNA Artificial Sequence Description of ArtificialSequencePrimer 10 taatacgact cactataggg 20 11 23 DNA Artificial SequenceDescription of Artificial SequencePrimer 11 actcactata gggctcgagc ggc 2312 22 DNA Artificial Sequence Description of Artificial SequencePrimer12 cacggaatcc tcagactctc ag 22 13 26 DNA Artificial Sequence Descriptionof Artificial SequencePrimer 13 gttgtagtac cacgctgatt ctctcc 26 14 63DNA Artificial Sequence Description of Artificial SequencePrimer 14gttctagagg aagatgaaga ggtcacggag gaggcagaaa tggagccaga agataaggga 60 cat63

1. A nucleic acid molecule comprising a nucleotide sequence encoding atubulin molecule, wherein said nucleic acid molecule comprises thesequence according to SEQ. ID. No. 1
 2. A nucleic acid moleculecomprising a nucleotide sequence encoding a tubulin molecule, whereinsaid nucleic acid molecule comprises the sequence according to SEQ. ID.No.
 2. 3. A nucleic acid molecule comprising a nucleotide sequenceencoding the C terminal end of a tubulin molecule, wherein the Cterminal end comprises the following sequence: VLEEDEEVTEEAEMEPEDKGH(SEQ ID. No. 4)
 4. A nucleic acid molecule comprising a nucleotidesequence encoding a tubulin molecule, wherein said nucleic acid moleculehybridizes to the nucleic acid molecule selected from the groupcomprising SEQ. ID. ID. No. 1, SEQ. ID. No 2, SEQ. ID. No. 4 andderivatives therefrom.
 5. A nucleic acid molecule comprising anucleotide sequence encoding a tubulin molecule, wherein the tubulinmolecule or part thereof comprises an amino acid sequence according toSEQ. ID. No.
 3. 6. A nucleic acid molecule comprising a nucleotidesequence encoding a tubulin molecule, wherein the nucleotide nucleicacid molecule would hybridize, but for the degeneracy of the geneticcode, to the nucleic acid molecule and/or the nucleotide sequenceaccording to any of claims 1 to
 5. 7. The nucleic acid moleculeaccording to any of claims 1 to 6, wherein the tubulin molecule is thetubulin molecule expressed in haematopoietic tissue.
 8. The nucleic acidmolecule according to any of claims 1 to 7, wherein the tubulin moleculeis β-tubulin, preferably an isotype class VI beta-tubulin, mostpreferably a human homologue of the vertebrate beta tubulin isotypeclass VI.
 9. The nucleic acid molecule according to any of claims 1 to8, wherein the nucleic acid molecule is an isolated nucleic acidmolecule.
 10. A vector comprising the nucleic acid molecule according toany of claims 1 to
 9. 11. The vector according to claim 10, wherein thevector is an expression vector.
 12. A cell, preferably a host cell,comprising the vector according to any of claims 10 or
 11. 13. An aminoacid sequence of a tubulin molecule, preferably of a beta-tubulinmolecule and more preferably of a class VI beta-tubulin, wherein theamino acid sequence is encoded by a nucleic acid molecule according toany of claims 1 to 9 or part thereof.
 14. A method for the production ofa tubulin molecule, preferably of a beta-tubulin molecule and morepreferably of a class VI beta-tubulin, characterized by culturing thecell according to claim 12 and isolating the protein product.
 15. Amethod for diagnosing polymorphism in a tubulin molecule, preferablyβ-tubulin, and more preferably an isotype class VI beta-tubulin, mostpreferably a human homologue of the vertebrate beta tubulin isotypeclass VI or in the nucleic acid coding such tubulin, wherein a nucleicacid according to any of claims 1 to 9 is used.
 16. Use of a nucleicacid according to any of claims 1 to 9 for diagnosis.
 17. A method forthe screening of anticancer drugs using a nucleic acid according to anyof claims 1 to 9 or any transcription and/or translation productthereof.
 18. Use of a nucleic acid according to any of claims 1 to 9 forscreening purposes.
 19. Use of a nucleic acid according to any of claims1 to 9 as a medicament.